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Specification Sheets
Safety Data Sheet
Intended UseFor Mohs In Vitro Diagnostic Use
Summary and Explanation

The Mohs Mouse/Rabbit PolyDetector DAB HRP Brown Detection System is a, 1-step Fab micropolymeric detection system that allows for the demonstration of antigens in cryostat sections, formalin-fixed paraffin-embedded tissue, and cell preparations. The Mohs PolyDetector kit has been developed using a micro-biopolymer conjugated to monomeric Fab’ immunoglobulin fractions targeting the Fc region of Mouse and Rabbit antibodies. Additionally, the micro-biopolymer is labeled with high quality HRP for maximum sensitivity. This ensures excellent cellular penetration which generates consistent, reproducible sensitive and specific immuno detection for all types of nuclear, cytoplasmic and membranous antigens, in different types of frozen, FFPE tissues and cell preparations.

The increased sensitivity of the Mohs Mouse/Rabbit PolyDetector DAB HRP Brown Detection System allows for rapid IHC procedures without compromising stain quality. The Mohs Mouse/Rabbit PolyDetector DAB HRP Brown Detection System is suitable for use with mouse IgG and IgM and rabbit primary antibodies, both monoclonal and polyclonal. The Mohs Mouse/Rabbit PolyDetector DAB HRP Brown Detection System kits are optimized for use with Bio SB TintoFast primary antibodies; however, they are universal kits and therefore should work equally well with antibodies from different vendors, as long as they are properly optimized.

PresentationThe Mohs Mouse/Rabbit PolyDetector DAB HRP Brown Detection System contains an anti-Mouse/Rabbit Horseradish Peroxidase micro-biopolymer solution, an HRP Brown Buffer Substrate, and a HRP Brown Chromogen solution. All components are buffered with stabilizers and a non-azide anti-microbial agent.
Availability
CAT.#PRESENTATIONVOL
BSB 0307SMohs Mouse/Rabbit PolyDetector DAB HRP Brown Detection System5.0 ml
BSB 0307Mohs Mouse/Rabbit PolyDetector DAB HRP Brown Detection System15.0 ml
BSB 0308Mohs Mouse/Rabbit PolyDetector DAB HRP Brown Detection System50.0 ml
BSB 0309Mohs Mouse/Rabbit PolyDetector DAB HRP Brown Detection System100.0 ml

Mohs IHC Procedure

Specimen Preparation of Mohs Frozen Tissues

  1. Embed the specimen in OCT inside a cryostat.
  2. Cut sections at 4-5 µm and mount on a positively charged glass slide such as the Bio SB Hydrophilic Plus Slides (BSB 7028) or the TintoDetector Cap Gap Plus (BSB 7006).
  3. Air dry the slide at room temperature for 2 minutes and then incubate the slide at 60 °C for 3 minutes in an incubator or dry bath.
  4. Fix in 100% acetone and air dry or fix with 10% NBF for 2 minutes at room temperature then rinse with distilled water and air dry.
  5. Frozen Tissue pretreatment depends on the target of interest. Refer to product insert form primary antibody to determine appropriate pretreatment protocol.

Tissue Pretreatment Procedure for Mohs Frozen Tissues

    1. Subject tissues to epitope retrieval using Bio SB ImmunoDNA Retriever with Citrate (BSB 0020-BSB 0023), ImmunoDNA Retriever with EDTA (BSB 0030-BSB 0033) or Mohs ImmunoDigestor (BSB 0324- 0326).  It is recommended that user choose either a 5 minutes heat-induced epitope retrieval (HIER) method using a pressure cooker (BSB TintoRetriever Pressure Cooker) at 110°C, or a 1-minute proteolytic induced epitope retrieval (PIER) method (for Cytokeratins using 10% NBF fixed tissues).  Morphology is better with 10% NBF fixed tissues, especially when using proteolytic induced epitope retrieval method, however, antigen maybe easier to detect on the 100% acetone fixed tissue.
    2. Any of three HIER methods may be used:

a. TintoRetriever Pressure Cooker or Equivalent

Place tissues/slides in a staining dish or coplin jar containing the ImmunoDNA Retriever with Citrate or EDTA, or TintoDeparaffinator Citrate or EDTA, and place on trivet in the pressure cooker. Add 1-2 inches of distilled water to the pressure cooker and turn heat to high (110-121° C). Incubate for 15 minutes. Release vapor, open and immediately transfer slides to room temperature.

b. TintoRetriever PT Module or Water Bath Method

Place tissues/slides in a pre-warmed staining dish or coplin jar containing the ImmunoDNA Retriever with Citrate or EDTA or TintoDeparaffinator Citrate or EDTA at 95°-99° C. Incubate for 30-60 minutes. Open immediatly and transfer slides to room temperature.

c. Conventional Steamer Method

Place tissues/slides in a pre-warmed staining dish or coplin jar containing the ImmunoDNA Retriever with Citrate or EDTA or Tintodeparaffinator Citrate or EDTA in a steamer, cover and steam for 30-60 minutes. After heat treatment, transfer slides in ImmunoDNA Retriever with Citrate or EDTA or in Tintodeparaffinator Citrate or EDTA to room temperature and let stand for 15-20 minutes.

3. For PIER, apply Mohs ImmunoDigestor (BSB 0324- 0326) at room temperature for 1 min then wash.

IHC Detection Procedure

  1. After HIER or PIER, transfer slides to ImmunoDNA washer and let stand for 1-2 minutes.
  2. For manual staining, perform antibody incubation at ambient temperature. For automated staining methods, perform antibody incubation according to instrument manufacturer’s instructions.
  3. Wash slides with ImmunoDNA washer or DI water.
  4. Continue Mohs IHC detection protocol. Wash slides between each step with ImmunoDNA washer solution.

Abbreviated Mohs Immunohistochemical Protocol

StepMohs PolyDetector HRP Brown Protocol
Primary Antibody4 min
Detection Step3 min
Substrate-Chromogen2 min
Counterstain / Coverslip0.5 min